library(tidyverse)
library(vcfR)
library(stringr)
library(RcppRoll)
cnames <- str_split("CHROM POS ID REF ALT QUAL FILTER INFO FORMAT S95 S96",pattern=boundary("word"))[[1]]
data <- read_tsv("S95S96_filtered.vcf.recode.vcf",comment="#",col_names = cnames)
Parsed with column specification:
cols(
CHROM = col_character(),
POS = col_integer(),
ID = col_character(),
REF = col_character(),
ALT = col_character(),
QUAL = col_double(),
FILTER = col_character(),
INFO = col_character(),
FORMAT = col_character(),
S95 = col_character(),
S96 = col_character()
)
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data
format.names <- str_split(data$FORMAT[[1]],pattern=":")[[1]]
data2 <- data %>%
select(-INFO,-ID,-FILTER, -FORMAT) %>%
mutate(S95=str_replace_all(S95,"([^0-9]|$)\\.","\\1NA")) %>% # convert missing data "." into NA
mutate(S96=str_replace_all(S96,"([^0-9]|$)\\.","\\1NA")) %>%
separate(S95,sep=":",into = str_c("S95_",format.names), convert = TRUE) %>%
separate(S96,sep=":",into=str_c("S96_",format.names), convert = TRUE) %>%
filter(!(S95_GT=="1/1" & S96_GT=="1/1")) %>%
filter(!(S95_GT=="0/0" & S96_GT=="0/0")) %>%
select(-ends_with("DPR"))
data2
data2 <- data2 %>%
mutate(S95_alt_AF=S95_AO/S95_DP,
S96_alt_AF=S96_AO/S96_DP,
S95_alt_major=S95_alt_AF > 0.5)
data2 <- data2 %>%
mutate(S95_AF_polar=ifelse(S95_alt_major,S95_alt_AF,1-S95_alt_AF),
S96_AF_polar=ifelse(S95_alt_major,S96_alt_AF,1-S96_alt_AF))
data2
to create a series of average values of different subsets of the full data set. a moving average can smooth out the noise of random outliers and emphasize long-term trends.
pl.data2 <- data2 %>%
gather(key="population",value="allele.frequency",S95_AF_polar,S96_AF_polar) %>%
arrange(CHROM,POS) %>%
group_by(population,CHROM) %>%
mutate(AF_100=roll_mean(allele.frequency,n=100,fill=NaN))
pl.data2
plot!
plA.data <- pl.data2 %>% filter(str_detect(CHROM,"chrA"),!str_detect(CHROM,"random"))
plA <- ggplot(plA.data,aes(x=POS,y=AF_100,color=population,shape=population)) +
facet_wrap(~CHROM, scales = "free_x",ncol=2) +
#geom_point(alpha=.1) +
geom_line()
plA
plot!
plC.data <- pl.data2 %>% filter(str_detect(CHROM,"chrC"),!str_detect(CHROM,"random"))
plC <- ggplot(plC.data,aes(x=POS,y=AF_100,color=population,shape=population)) +
facet_wrap(~CHROM, scales = "free_x",ncol=2) +
#geom_point(alpha=.1) +
geom_line()
plC
plot one per chromosome
chrom.names <- unique(pl.data2$CHROM) %>% str_subset("[0-9]$")
chrom.names
for (chr in chrom.names) {
pl.data2 %>% filter(str_detect(CHROM,chr)) %>%
ggplot(aes(x=POS,y=AF_100,color=population)) +
geom_line() +
ggtitle(chr) +
ylim(0,1)
ggsave(str_c(chr,".png"),height = 8, width = 12)
}
data3 <- data2 %>%
group_by(CHROM) %>%
mutate(abs_delta_af = abs(S95_AO/S95_DP - S96_AO/S96_DP),
abs_delta_af_100=roll_mean(abs_delta_af,n=100,fill=NaN),
abs_delta_af_500=roll_mean(abs_delta_af,n=500,fill=NaN)
)
data3
plot one per chromosome
for (chr in chrom.names) {
data3 %>% filter(str_detect(CHROM,chr)) %>%
ggplot(aes(x=POS,y=abs_delta_af_100)) +
geom_line() +
ggtitle(chr) +
ylim(0,1)
ggsave(str_c("Delta_af",chr,".png"),height = 8, width = 12)
}
ggplot(data3, aes(x=S95_DP)) + geom_histogram()
ggplot(data3, aes(x=S96_DP)) + geom_histogram()
F2snps <- read_tsv("Final_F2_SNP_Sites.tab")
data2.F2.filtered <- semi_join(data2,F2snps)
dim(data2)
[1] 1926088 26
dim(F2snps)
[1] 18226 2
dim(data2.F2.filtered)
[1] 1997 26
data3.F2 <- data2.F2.filtered %>%
group_by(CHROM) %>%
mutate(abs_delta_af = abs(S95_AO/S95_DP - S96_AO/S96_DP),
abs_delta_af_10=roll_mean(abs_delta_af,n=10,fill=NaN)
)
data3.F2
for (chr in chrom.names) {
data3.F2 %>% filter(str_detect(CHROM,chr)) %>%
ggplot(aes(x=POS,y=abs_delta_af_10)) +
geom_line() +
ggtitle(chr) +
ylim(0,1)
ggsave(str_c("Delta_af_F2_filter",chr,".png"),height = 8, width = 12)
}
snps.505 <- read_csv("505_Genotype_170K_SNPs.csv.gz") %>%
select(CHROM,POS)
head(snps.505)
data2.505.filtered <- semi_join(data2,snps.505) #return all rows from data2 where there are matching values in snp.505
Joining, by = c("CHROM", "POS")
dim(data2)
[1] 1926088 26
dim(snps.505)
[1] 174397 2
dim(data2.505.filtered)
[1] 14116 26
data3.505 <- data2.505.filtered %>%
group_by(CHROM) %>%
mutate(abs_delta_af = abs(S95_AO/S95_DP - S96_AO/S96_DP),
abs_delta_af_10=roll_mean(abs_delta_af,n=10,fill=NaN)
)
data3.505
data3.505 %>% filter(!grepl("random", CHROM)) %>%
ggplot(aes(x=POS,y=abs_delta_af_10,group=CHROM,color=CHROM)) +
geom_point() +
geom_line() +
ggtitle("absolute diff. of allele freq. between Rf and MS pool") +
ylim(0,1)
ggsave("Delta_af_505_filter_entire_genome.png", width = 16, height = 20, device = "png")
We pick chr09 and will focus on it!
for (chr in chrom.names) {
data3.505 %>% filter(str_detect(CHROM,chr)) %>%
ggplot(aes(x=POS,y=abs_delta_af_10)) +
geom_point(aes(y=abs_delta_af),shape=1) +
geom_line(color="blue") +
ggtitle(chr) +
ylim(0,1)
ggsave(str_c("Delta_af_505_filter",chr,".png"),height = 8, width = 12)
}
library(rtracklayer)
library(IRanges)
library(stringr)
genes <- read.delim("Brassica_napus.annotation_v5_modified_modified.gff3", sep="\t", comment.char="#", as.is=T, header=F)
head(genes)
colnames(genes) <- c("chromosome","source","type","start","end","score","strand","phase","attributes")
head(genes)
genes <- genes %>% filter(type=="mRNA")
genes$locus <- sub(pattern = "ID=", "", sub(pattern = ";.*", replacement = "", x = genes$attributes))
head(genes)
genes.rd <- RangedData(IRanges(start=genes$start, end=genes$end,names=genes$locus),space=genes$chromosome)
head(genes.rd)
RangedData with 6 rows and 0 value columns across 41 spaces
space ranges |
<factor> <IRanges> |
BnaA01g23190D chrA01 [15588360, 15589835] |
BnaA01g23200D chrA01 [15603637, 15608043] |
BnaA01g23210D chrA01 [15616241, 15617098] |
BnaA01g23220D chrA01 [15617294, 15619886] |
BnaA01g23230D chrA01 [15634005, 15635182] |
BnaA01g23240D chrA01 [15647464, 15648070] |
region1.rd <- RangedData(IRanges(start=4.371e07,end=4.372e07,names="Region1"),space="chrC09")
region1.rd
RangedData with 1 row and 0 value columns across 1 space
space ranges |
<factor> <IRanges> |
Region1 chrC09 [43710000, 43720000] |
overlaps1_4.371_4.372 <- as.data.frame(as.matrix(findOverlaps(ranges(region1.rd),ranges(genes.rd))))
nrow(overlaps1_4.371_4.372) #1,420 hits
[1] 4
overlaps1_4.371_4.372$locus <- genes$locus[overlaps1_4.371_4.372$subjectHits]
write.csv(overlaps1_4.371_4.372,"overlaps1_4.371_4.372.csv")
napus_vs_ara <- read.table("napus_vs_ara.non_reciprocal.table") #matches of gene functions between B.napus and Ara
position 4E7-5E7 on chrC09
overlaps1_4_5 <- read.csv("overlaps1_4_5.csv")
a <- overlaps1_4_5 %>%
mutate(napus_ID=locus) %>% select(subjectHits, napus_ID)
overlaps1_4_5_match_ara <- semi_join(napus_vs_ara, a, by="napus_ID")
overlaps1_4_5_match_ara #934 hits
position 4.35E7-4.38E7 on chrC09
overlaps1_4.35_4.38 <- read.csv("overlaps1_4.35_4.38.csv")
b <- overlaps1_4.35_4.38 %>%
mutate(napus_ID=locus) %>% select(subjectHits, napus_ID)
overlaps1_4.35_4.38_match_ara <- semi_join(napus_vs_ara, b, by="napus_ID")
overlaps1_4.35_4.38_match_ara #24 hits
position 4.370E7-4.373E7 on chrC09
overlaps1_4.370_4.373 <- read.csv("overlaps1_4.370_4.373.csv")
b <- overlaps1_4.370_4.373 %>%
mutate(napus_ID=locus) %>% select(subjectHits, napus_ID)
overlaps1_4.370_4.373_match_ara <- semi_join(napus_vs_ara, b, by="napus_ID")
overlaps1_4.370_4.373_match_ara #1 hit
[BnaC08g07750D AT2G38670.1 phosphorylethanolamine cytidylyltransferase 1] (https://www.arabidopsis.org/servlets/TairObject?id=35231&type=locus)